Expression and characterization of extreme alkaline, oxidation-resistant keratinase from Bacillus licheniformis in recombinant Bacillus subtilis WB600 expression system and its application in wool fiber processing

2012 ◽  
Vol 29 (5) ◽  
pp. 825-832 ◽  
Author(s):  
Baihong liu ◽  
Juan Zhang ◽  
Ben Li ◽  
Xiangru Liao ◽  
Guocheng Du ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Licheniformis ◽  
Expression System ◽  
Wool Fiber ◽  
Oxidation Resistant ◽  
Recombinant Bacillus Subtilis ◽  
Fiber Processing ◽  
Alkaline Oxidation ◽  
Bacillus Subtilis Wb600

scholarly journals Autolysis of isolated cell walls of Bacillus licheniformis N.C.T.C. 6346 and Bacillus subtilis Marburg strain 168. Separation of the products and characterization of the mucopeptide fragments

1970 ◽  
Vol 119 (5) ◽  
pp. 849-860 ◽  
Author(s):  
R. C. Hughes
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Licheniformis ◽  
Cell Walls ◽  
Deae Cellulose ◽  
Casein Hydrolysate ◽  
Diaminopimelic Acid ◽  
Average Chain Length ◽  
Amide Bonds ◽  
Chain Lengths

1. Cell walls were isolated from Bacillus licheniformis N.C.T.C. 6346 and Bacillus subtilis Marburg strain 168 trp grown on casein hydrolysate into exponential phase. Autolysis was carried out and the soluble products, separated by chromatography on DEAE-cellulose, from the two wall preparations are broadly similar in composition and are in agreement with autolysis proceeding with hydrolysis of amide bonds between l-alanine and N-acetylmuramic acid residues in the mucopeptide components. 2. Peptides originating from the mucopeptide components were isolated and shown to be a monomer peptide, l-alanyl-d-glutamyl-meso-diaminopimelic acid and a dimer peptide containing two monomer peptides linked through a residue of d-alanine. Approximately one amide group is present for each equivalent tripeptide unit and is probably substituted on diaminopimelic acid residues. 3. Oligosaccharides originating from the mucopeptide components were isolated and after hydrolysis contained almost equimolar amounts of glucosamine and muramic acid and only very small amounts of amino acids. The number-average chain length, estimated by the release of non-reducing end groups of N-acetylglucosamine with exo-β-N-acetylglucosaminidase, is approximately ten hexosamine residues for oligosaccharides isolated from either organism. The oligosaccharides are polydisperse. 4. N-Acetylglucosamine residues are the only reducing terminals detectable in the oligosaccharides isolated from B. subtilis or B. licheniformis cell-wall autolysates. The number-average chain lengths of the oligosaccharides were determined by estimation of the content of these residues and are higher than those found by enzymic assay. Possible reasons for the discrepancy are discussed.

scholarly journals Characterization of the sacQ genes from Bacillus licheniformis and Bacillus subtilis.

1987 ◽  
Vol 169 (1) ◽  
pp. 324-333 ◽  
Author(s):  
A Amory ◽  
F Kunst ◽  
E Aubert ◽  
A Klier ◽  
G Rapoport
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Licheniformis

scholarly journals Development and Characterization of a Subtilin-Regulated Expression System in Bacillus subtilis: Strict Control of Gene Expression by Addition of Subtilin

2005 ◽  
Vol 71 (12) ◽  
pp. 8818-8824 ◽  
Author(s):  
Roger S. Bongers ◽  
Jan-Willem Veening ◽  
Maarten Van Wieringen ◽  
Oscar P. Kuipers ◽  
Michiel Kleerebezem
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Fluorescent Protein ◽  
Expression System ◽  
Functional Characterization ◽  
Industrial Applications ◽  
Wild Type ◽  
Control Of Gene Expression ◽  
Regulated Gene Expression

ABSTRACT A system for subtilin-regulated gene expression (SURE) in Bacillus subtilis that is based on the regulatory module involved in cell-density-dependent control of the production of subtilin is described. An integration vector for introduction of the essential sensor-regulator couple spaRK into the amyE locus of the B. subtilis chromosome and a B. subtilis 168-derived production host in which the spaRK genes were functionally introduced were constructed. Furthermore, several expression plasmids harboring the subtilin-inducible wild-type spaS promoter or a mutated derivative of this promoter were constructed, which facilitated both transcriptional and translational promoter-gene fusions. Functional characterization of both spaS promoters and the cognate expression host could be performed by controlled overproduction of the β-glucuronidase (GusA) and green fluorescent protein (GFP) reporters. Both spaS promoters exhibited very low levels of basal expression, while extremely high levels of expression were observed upon induction with subtilin. Moreover, the level of expression depended directly on the amount of inducer (subtilin) used. The wild-type spaS promoter appeared to be more strictly controlled by the addition of subtilin, while the highest levels of expression were obtained when the mutated spaS promoter was used. Induction by subtilin led to 110- and 80-fold increases in GusA activity for the spaS promoter and its mutant derivative, respectively. Since the SURE system has attractive functional characteristics, including promoter silence under noninducing conditions and a controlled and high level of expression upon induction, and since it is not subject to catabolite control, we anticipate that it can provide a suitable expression system for various scientific and industrial applications.

scholarly journals Overexpression of the Bacillus licheniformis GroES enhances thermotolerance of Bacillus subtilis WB600

2018 ◽  
Vol 32 (6) ◽  
pp. 1527-1532 ◽  
Author(s):  
Zixing Dong ◽  
Xiaoling Chen ◽  
Ke Cai ◽  
Peili Shen ◽  
Kangming Tian ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Licheniformis ◽  
Bacillus Subtilis Wb600

Recombinant Expression and Biochemical Characterization of Levansucrase from Halophilic Bacteria Bacillus licheniformis BK1 and BK2

2021 ◽  
Vol 874 ◽  
pp. 96-106
Author(s):  
Nur Umriani Permatasari ◽  
Enny Ratnaningsih ◽  
Rukman Hertadi
Keyword(s):  
Bacillus Licheniformis ◽  
Biochemical Characterization ◽  
Specific Activity ◽  
Expression System ◽  
Halophilic Bacteria ◽  
Optimum Conditions ◽  
E Coli ◽  
Optimization Result ◽  
Rsm Optimization

Levansucrase was an extracellular polysacharride (EPS) which has a role in synthesizing levans by transferring fructose moiety from sucrose to acceptor molecules. In the previous study, we have successfully cloned the levansucarese gene from two Bacillus licheniformis strains of BK1 and BK2 labeled as lsbl-bk1 and lsbl-bk2. The present study aims to optimize the expression level of both genes in E. coli expression system and also to obtain the optimum conditions for the recombinant enzymes activity by applying the response surface methodology (RSM). The optimization result found that the highest Lsbl-bk1 production in E. coli expression system was occurred when the recombinant cells grown in the medium containing 0.6% (w/v) NaCl at 42°C, and induced by 0.6 mM IPTG. Different optimum conditions were found for Lsbl-bk2 production. It was achieved when 1.1% (w/v) NaCl added to the production medium and induced by 0.7 mM IPTG at 40°C. RSM optimization result for biochemical characterization of Lsbl-bk1 levansucrase showed the highest specific activity achieved at 56°C and pH 7.5, whereas for the Lsbl-bk2 levansucrase reached the highest specific activity at 50°C and pH 7.5. The addition of Co2+, Ti2+, Mg2+, Ba2+, Zn2+, Fe3+, Ca2+ metal ion to both levansucrases solution did not significantly altered their specific activity, indicating that both levansucrases are not metalo enzymes. Furthermore, the specific activity of levansucrase was also not affected by the addition of 1-25% (w/v) NaCl, suggesting that the variation of ionic strength did not alter the native state of both enzymes. The plot results of levansucrase specific activities toward sucrose concentration showed that both levansucrases follow Michaelis-Menten profile with kcat/KM values ​​about 3.8 and 3.6 s-1/mM respectively. These data indicated that the recombinant levansucrases from halophilic bacteria B. licheniformis BK1 and BK2 are a non metaloenzyme with high affinity and binding rate to sucrose substrate, in which the catalytic efficiency on hydrolysis reactions is relatively low.

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